June 2017
Volume 58, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2017
Patch clamp recordings from mouse cone photoreceptores
Author Affiliations & Notes
  • Norianne Theresa Ingram
    Integrative Biology and Physiology, UCLA, Los Angeles, California, United States
    Ophthalmology, The Jules Stein Institute, UCLA, Los Angeles, California, United States
  • Gordon Fain
    Integrative Biology and Physiology, UCLA, Los Angeles, California, United States
    Ophthalmology, The Jules Stein Institute, UCLA, Los Angeles, California, United States
  • Alapakkam P Sampath
    Ophthalmology, The Jules Stein Institute, UCLA, Los Angeles, California, United States
  • Footnotes
    Commercial Relationships   Norianne Ingram, None; Gordon Fain, None; Alapakkam Sampath, None
  • Footnotes
    Support  EY001844, EY17606
Investigative Ophthalmology & Visual Science June 2017, Vol.58, 1024. doi:
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      Norianne Theresa Ingram, Gordon Fain, Alapakkam P Sampath; Patch clamp recordings from mouse cone photoreceptores. Invest. Ophthalmol. Vis. Sci. 2017;58(8):1024.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose : The difficulty of making physiological and biochemical measurements from mouse cones has proved a substantial hurdle to understanding their function; as a result, much more is known about mammalian rods than cones. We are able to identify and patch-clamp single, unlabeled mouse cones even though cones represent only 3% of mouse photoreceptors. Voltage-clamp recording gives tight control over the electronics of the clamped cell and has not been used to study mouse cone phototransduction.

Methods : Mouse retinas were isolated under infrared illumination. Slices were cut to maintain vertical circuitry. Recordings were made from individual cone somas recorded in voltage-clamp or current-clamp in response to 405 nm light, which stimulates both spectral classes of cones equally.

Results : We present the first voltage and current recordings from the same mouse cone. The maximum photocurrent averaged 24.5 ± 2.7 pA. (SEM, n =18) Wild-type (WT) cone flash and step responses show variable evidence of rod input, while Gnat1-/- cones do not show rod tails. We will also show flash and step responses of GCAP-/- and Rv-/- cones. A comparison of current and voltage responses in rods reveals significant temporal filtering: the photovoltage peaks before photocurrent and decays more rapidly. We do not see the same photovoltage speeding for mouse cones.

Conclusions : Individual mouse cones can be reliably located and patched in WT and other transgenic lines. Responses are 3-4 times larger than in previous studies using suction electrodes, yielding a greater signal-to-noise ratio. As the slice preparation maintains functional circuitry, our method can also be used to investigate the electrical properties of cones on a network level in addition to single-cell analyses. We propose this method as a powerful technique for probing cone physiology.

This is an abstract that was submitted for the 2017 ARVO Annual Meeting, held in Baltimore, MD, May 7-11, 2017.

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